In vivo profiling of DPP4 inhibitors reveals alterations in collagen metabolism and accumulation of an amyloid peptide in rat plasma.
Dipeptidyl peptidase 4 (DPP4) inhibitors represent a novel class of oral anti-hyperglycemic agents. The complete pharmacological profile of these protease inhibitors remains unclear. In order to gain deeper insight into the in vivo effects caused by DPP4 inhibition, two different DPP4 inhibitors (vildagliptin and AB192) were analyzed using differential peptide display. Wistar rats were treated with the DPP4 inhibitors (0.3mgkg(-1); 1mgkg(-1) or 3mgkg(-1) body weight) and DPP4 activity was measured before and at the end of the experiment. One hour after compound administration, blood plasma samples were collected to generate peptide displays and to subsequently identify differentially regulated peptides. A dose-dependent decrease in blood plasma DPP4 activity was measured for both inhibitors. DPP4 inhibition influenced collagen metabolism leading to depletion of collagen derived peptides (e.g. collagen alpha 1 (III) 521-554) and accumulation of related N-terminally extended collagen derived peptides (e.g. collagen alpha 1 (III) 519-554). Furthermore, the intact amyloid rat BRI (1-23) peptide was detected in plasma following in vivo DPP4 inhibition. DPP4 catalyzed cleavage kinetics of the BRI peptide were determined in vitro. The k(cat) and K(m) for cleavage by DPP4 were 5.2s(-1) and 14microM, respectively, resulting in a specificity constant k(cat)/K(m) of 0.36 x 10(6)s(-1)M(-1). Our results demonstrate that differential peptide analysis can be applied to monitor action of DPP4 inhibition in blood plasma. For the first time effects on basal collagen metabolism following DPP4 inhibition in vivo were demonstrated and the BRI amyloid peptide was identified as a novel DPP4 substrate.
Jost
Marco M
MM
Digilab BioVisioN GmbH, Hannover, Germany.
Lamerz
Jens
J
Tammen
Harald
H
Menzel
Christoph
C
De Meester
Ingrid
I
Lambeir
Anne-Marie
AM
Augustyns
Koen
K
Scharpé
Simon
S
Zucht
Hans Dieter
HD
Rose
Horst
H
Jürgens
Michael
M
Schulz-Knappe
Peter
P
Budde
Petra
P
D000218:Adamantane / Q000031:analogs & derivatives* / Q000494:pharmacology; D000682:Amyloid / Q000097:blood*; D000818:Animals; D003094:Collagen / Q000378:metabolism*; D018819:Dipeptidyl Peptidase 4 / Q000097:blood* / Q000737:chemistry / Q000302:isolation & purification; D054873:Dipeptidyl-Peptidase IV Inhibitors / Q000494:pharmacology*; D006352:Heart Ventricles / Q000201:enzymology; D006801:Humans; D007700:Kinetics; D009570:Nitriles / Q000494:pharmacology*; D063065:Organophosphonates / Q000494:pharmacology*; D010446:Peptide Fragments / Q000097:blood* / Q000302:isolation & purification; D011392:Proline / Q000031:analogs & derivatives* / Q000494:pharmacology; D011759:Pyrrolidines / Q000494:pharmacology*; D051381:Rats; D017208:Rats, Wistar; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013379:Substrate Specificity; D000077597:Vildagliptin
D000682:Amyloid; D054873:Dipeptidyl-Peptidase IV Inhibitors; D009570:Nitriles; D063065:Organophosphonates; D010446:Peptide Fragments; D011759:Pyrrolidines; C118220:bis(4-acetamidophenyl) 1-prolylpyrrolidine-2-phosphonate; D003094:Collagen; D011392:Proline; D018819:Dipeptidyl Peptidase 4; D000077597:Vildagliptin; D000218:Adamantane